Substrate-bound outward-open state of the betaine transporter BetP provides insights into Na+ coupling.

The Na(+)-coupled betaine symporter BetP shares a highly conserved fold with other sequence unrelated secondary transporters, for example, with neurotransmitter symporters. Recently, we obtained atomic structures of BetP in distinct conformational states, which elucidated parts of its alternating-access mechanism. Here, we report a structure of BetP in a new outward-open state in complex with an anomalous scattering substrate, adding a fundamental piece to an unprecedented set of structural snapshots for a secondary transporter.

Phlorins bearing different substituents at the sp3-hybridized meso-position.

Phlorins bearing different substituents at the sp(3)-hybridized meso-position were investigated. The extent to which different substituents at this unique position can influence phlorin spectroscopic properties, structure, and stability is of interest given that such substituents are not in direct conjugation with the phlorin macrocycle. While the effect of various substituents at the sp(2)-hybridized positions has been the subject of prior investigations, the impact of different substituents at the saturated carbon atom has not been systematically examined.

Facile solid phase peptide synthesis with a Re-lysine conjugate generated via a one-pot procedure.

We have synthesized a Re(CO)3-modified lysine via a one-pot Schiff base formation reaction that can be used in the solid phase peptide synthesis. To demonstrate its potential use, we have attached it to a neurotensin fragment and observed uptake into human umbilical vascular endothelial cells.

Structural insights into functional lipid-protein interactions in secondary transporters.

Background: Structural evidences with functional corroborations have revealed distinct features of lipid-protein interactions especially in channels and receptors. Many membrane embedded transporters are also known to require specific lipids for their functions and for some of them cellular and biochemical data suggest tight regulation by the lipid bilayer. However, molecular details on lipid-protein interactions in transporters are sparse since lipids are either depleted from the detergent solubilized transporters in three-dimensional crystals or not readily resolved in crystal structures.

Computational algorithm-driven evaluation of monocytic myeloid-derived suppressor cell frequency for prediction of clinical outcomes.

Evaluation of myeloid-derived suppressor cells (MDSC), a cell type implicated in T-cell suppression, may inform immune status. However, a uniform methodology is necessary for prospective testing as a biomarker. We report the use of a computational algorithm-driven analysis of whole blood and cryopreserved samples for monocytic MDSC (m-MDSC) quantity that removes variables related to blood processing and user definitions.

Bystander chronic infection negatively impacts development of CD8(+) T cell memory.

Epidemiological evidence suggests that chronic infections impair immune responses to unrelated pathogens and vaccines. The underlying mechanisms, however, are unclear and distinguishing effects on priming versus development of immunological memory has been challenging. We investigated whether bystander chronic infections impact differentiation of memory CD8(+) T cells, the hallmark of protective immunity against intracellular pathogens.

A convenient, optimized pipeline for isolation, fluorescence microscopy and molecular analysis of live single cells.

Background: Heterogeneity within cell populations is relevant to the onset and progression of disease, as well as development and maintenance of homeostasis. Analysis and understanding of the roles of heterogeneity in biological systems require methods and technologies that are capable of single cell resolution. Single cell gene expression analysis by RT-qPCR is an established technique for identifying transcriptomic heterogeneity in cellular populations, but it generally requires specialized equipment or tedious manipulations for cell isolation.

Mutation of a single threonine in the cytoplasmic NH2 terminus disrupts trafficking of renal betaine-GABA transporter 1 during hypertonic stress.

Betaine is an important osmolyte and is, compared with other organs, much more abundant in the kidneys, where it enters cells in the medulla by betaine-GABA transporter 1 (BGT1) to balance osmoregulation in the countercurrent system. In wild-type (wt-)BGT1-expressing oocytes, GABA-mediated currents were diminished by preincubation of oocytes with 100 nM PMA or 5 μM dioctanoyl-sn-glycerol, activators of PKC, whereas the application of staurosporine before the application of dioctanoyl-sn-glycerol restored the response to GABA. Four potential phosphorylation sites on BGT1 were mutated to alanine by site-directed mutagenesis.

Pharmacoepigenetics of depression: no major influence of MAO-A DNA methylation on treatment response.

The monoamine oxidase A (MAO-A) gene has been suggested to be involved in the pathogenesis as well as the pharmacological treatment of major depressive disorder. In the present analysis, for the first time a pharmacoepigenetic approach was applied investigating the influence of DNA methylation patterns in the MAO-A regulatory and exon1/intron1 region on antidepressant treatment response. 94 patients of Caucasian descent with major depressive disorder (f = 61; DSM-IV) were analyzed for DNA methylation status at 43 MAO-A CpG sites via direct sequencing of sodium bisulfite treated DNA extracted from blood cells.

Synthesis, redox properties, and electronic coupling in the diferrocene aza-dipyrromethene and azaBODIPY donor-acceptor dyad with direct ferrocene-α-pyrrole bond.

3,3'-Diferrocenylazadipyrromethene (3) and corresponding difluoroboryl (azaBODIPY) complex (4) were synthesized in several steps from ferrocenecarbaldehyde, following the well-explored chalcone-type synthetic approach. The novel diiron complexes, in which ferrocene groups are directly connected to the α-pyrrolic positions were characterized by a variety of spectroscopic techniques, electrochemistry, spectroelectrochemistry, and X-ray crystallography, while their electronic structure, redox properties, and UV-vis spectra were correlated with the density functional theory (DFT) and time-dependent DFT calculations.

Rabbit anti-T-lymphocyte globulin (ATG) persists with differential reactivity in patients' sera after full hematopoetic regeneration from allogeneic stem cell transplantation.

Background: Rabbit polyclonal anti-T-lymphocyte Globulin (ATG-F®, Fresenius) is widely used for GvHD prophylaxis in allogeneic stem cell transplantation (SCT). ATG has a wide epitope spectrum and has been shown to react with all compartments of peripheral blood mononuclear cells (PBMNCs). ATG induces apoptosis in all cellular compartments.

A new highly fluorescent and symmetric pyrrole-BF2 chromophore: BOPHY.

The new fluorescent chromophore BOPHY can be readily synthesized in two steps from commercially available reagents via the coupling of pyrrole-2-carboxaldehyde with hydrazine followed by reaction with BF3. The resultant symmetric and dimeric tetracycle is composed of two BF2 units in six-membered chelate rings appended with pyrrole units on the periphery. The quantum yields of fluorescence for the unmodified compound and the tetramethyl variant are near unity, with values of 95 and 92%, respectively, in CH2Cl2.

New diamino-diheterophenol ligands coordinate iron(III) to make structural and functional models of protocatechuate 3,4-dioxygenase.

Three diamino, dihetero-phenol ligands were synthesized by sequential Mannich condensations. These ligands were combined with FeCl3 to produce three five-coordinate Fe(III) complexes that are structural models for the enzyme 3,4-PCD. The three Fe(III) complexes were characterized by elemental analysis, single crystal X-ray diffraction studies, UV-vis spectroscopy, and cyclic voltammetry.

Adrenaline for out-of-hospital cardiac arrest resuscitation: a systematic review and meta-analysis of randomized controlled trials.

Introduction: The evidence for adrenaline in out-of-hospital cardiac arrest (OHCA) resuscitation is inconclusive. We systematically reviewed the efficacy of adrenaline for adult OHCA.

Methods: We searched in MEDLINE, EMBASE, and Cochrane Library from inception to July 2013 for randomized controlled trials (RCTs) evaluating standard dose adrenaline (SDA) to placebo, high dose adrenaline (HDA), or vasopressin (alone or combination) in adult OHCA patients.

Acetabular cartilage assessment in patients with femoroacetabular impingement by using T2* mapping with arthroscopic verification.

Purpose: To evaluate the ability of T2* mapping to help differentiate damaged from normal acetabular cartilage in patients with femoroacetabular impingement (FAI).

Materials And Methods: The institutional review board approved this retrospective study, and the requirement to obtain informed consent was waived. The study complied with HIPAA guidelines.

Multidimensional single-cell analysis of BCR signaling reveals proximal activation defect as a hallmark of chronic lymphocytic leukemia B cells.

Purpose: Chronic Lymphocytic Leukemia (CLL) is defined by a perturbed B-cell receptor-mediated signaling machinery. We aimed to model differential signaling behavior between B cells from CLL and healthy individuals to pinpoint modes of dysregulation.

Experimental Design: We developed an experimental methodology combining immunophenotyping, multiplexed phosphospecific flow cytometry, and multifactorial statistical modeling.

Incidence, indications, and predictors of adverse outcomes of postpartum hysterectomies: 20-year experience in a tertiary care centre.

Objective: Postpartum hysterectomy is an uncommon yet serious obstetric procedure associated with maternal morbidity and mortality. The objectives of our study were to assess the incidence of and indications for PH and to identify predictors of massive hemorrhage and coagulopathy.

Methods: We conducted a retrospective cohort study on all cases of PH performed at the Jewish General Hospital, McGill University, between 1992 and 2011.

Crystalline carbon nitride nanosheets for improved visible-light hydrogen evolution.

Nanosheets of a crystalline 2D carbon nitride were obtained by ionothermal synthesis of the layered bulk material poly(triazine imide), PTI, followed by one-step liquid exfoliation in water. Triazine-based nanosheets are 1-2 nm in height and afford chemically and colloidally stable suspensions under both basic and acidic conditions. We use solid-state NMR spectroscopy of isotopically enriched, restacked nanosheets as a tool to indirectly monitor the exfoliation process and carve out the chemical changes occurring upon exfoliation, as well as to determine the nanosheet thickness.

Sharing What We Know about Living a Good Life: Indigenous Approaches to Knowledge Translation.

Knowledge Translation (KT), a core priority in Canadian health research, policy, and practice for the past decade, has a long and rich tradition within Indigenous communities. In Indigenous knowledge systems the processes of "knowing" and "doing" are often intertwined and indistinguishable. However, dominant KT models in health science do not typically recognize Indigenous knowledge conceptualizations, sharing systems, or protocols and will likely fall short in Indigenous contexts.

Newly engineered cyan fluorescent proteins with enhanced performances for live cell FRET imaging.

Cyan fluorescent proteins (CFPs) derived from Aequorea victoria green fluorescent protein are the most widely used Förster resonant energy transfer (FRET) donors in genetically encoded biosensors for live-cell imaging and bioassays. However, the weak and complex fluorescence emission of cyan variants, such as enhanced cyan fluorescent protein (ECFP) or Cerulean, has long remained a major bottleneck in these FRET techniques. Recently, several CFPs with greatly improved performances, including mTurquoise, mTurquoise2, mCerulean3, and Aquamarine, have been engineered through a mixture of site-directed and large-scale random mutagenesis.

Thymic stromal lymphopoietin-mediated extramedullary hematopoiesis promotes allergic inflammation.

Extramedullary hematopoiesis (EMH) refers to the differentiation of hematopoietic stem cells (HSCs) into effector cells that occurs in compartments outside of the bone marrow. Previous studies linked pattern-recognition receptor (PRR)-expressing HSCs, EMH, and immune responses to microbial stimuli. However, whether EMH operates in broader immune contexts remains unknown.

Novel materials for biofilm reactors and their characterization.

The application of adherently growing microorganisms for biotechnological production processes is established, but it is still a niche technology with only a small economic impact. However, novel approaches are under development for new types of biofilm reactors. In this context, increasingly more microstructured metal surfaces are being investigated, and they show positive effects on the bacterial growth and the biofilm establishment.

Histone deacetylase 3 coordinates commensal-bacteria-dependent intestinal homeostasis.

The development and severity of inflammatory bowel diseases and other chronic inflammatory conditions can be influenced by host genetic and environmental factors, including signals derived from commensal bacteria. However, the mechanisms that integrate these diverse cues remain undefined. Here we demonstrate that mice with an intestinal epithelial cell (IEC)-specific deletion of the epigenome-modifying enzyme histone deacetylase 3 (HDAC3(ΔIEC) mice) exhibited extensive dysregulation of IEC-intrinsic gene expression, including decreased basal expression of genes associated with antimicrobial defence.

Structural evidence for functional lipid interactions in the betaine transporter BetP.

Bilayer lipids contribute to the stability of membrane transporters and are crucially involved in their proper functioning. However, the molecular knowledge of how surrounding lipids affect membrane transport is surprisingly limited and despite its general importance is rarely considered in the molecular description of a transport mechanism. One reason is that only few atomic resolution structures of channels or transporters reveal a functional interaction with lipids, which are difficult to detect in X-ray structures per se.

Magnetic circular dichroism spectroscopy of N-confused porphyrin and its ionized forms.

N-Confused porphyrin (NCP) and its externally methylated variant (MeNCP) were investigated using UV-vis and magnetic circular dichrosim (MCD) spectroscopies. In addition to evaluating the spectroscopy of the neutral compounds, the acid/base chemistry of these macrocycles was examined by the same methods. NCP exhibits two tautomeric states depending on the polarity of the solvent, and their protonation/deprotonation chemistries also differ depending on solvent polarity.